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The stress response of turbot Scophthalmus maximus was evaluated in fish maintained 8 days under different water depths, normal (NWD, 30 cm depth, total water volume 40 l) or low (LWD, 5 cm depth, total water volume 10 l), in the additional presence of infection–infestation of two pathogens of this species. This was caused by intraperitoneal injection of sublethal doses of the bacterium Aeromonas salmonicida subsp. salmonicida or the parasite Philasterides dicentrarchi (Ciliophora:Scuticociliatida). The LWD conditions were stressful for fish, causing increased levels of cortisol in plasma, decreased levels of glycogen in liver and nicotinamide adenine dinucleotide phosphate (NADP) and increased activities of G6Pase and GSase. The presence of bacteria or parasites in fish under NWD resulted in increased cortisol levels in plasma whereas in liver, changes were of minor importance including decreased levels of lactate and GSase activity. The simultaneous presence of bacteria and parasites in fish under NWD resulted a sharp increase in the levels of cortisol in plasma and decreased levels of glucose. Decreased levels of glycogen and lactate and activities of GSase and glutathione reductase (GR), as well as increased activities of glucose-6-phosphate dehydrogenase (G6PDH), 6-phosphogluconate dehydrogenase (6PGDH) and levels of nicotinamide adenine dinucleotide phosphate (NADPH) occurred in the same fish in liver. Finally, the presence of pathogens in S. maximus under stressful conditions elicited by LWD resulted in synergistic actions of both type of stressors in cortisol levels. In liver, the presence of bacteria or parasites induced a synergistic action on several variables such as decreased activities of G6Pase and GSase as well as increased levels of NADP and NADPH and increased activities of GPase, G6PDH and 6PGDH.

Down-regulation of fecundity through oocyte resorption was assessed in Baltic Sea turbot Scophthalmus maximus at three locations in the period from late vitellogenesis in April to spawning during June to July. The mean ± s.d. total length of the sampled fish was 32.7 ± 3.1 cm and mean ± s.d. age was 6.2 ± 1.5 years. Measurements of atresia were performed using the 'profile method' with the intensity of atresia adjusted according to the 'dissector method' (10.6% adjustment; coefficient of determination was 0.675 between methods). Both prevalence (portion of fish with atresia) and intensity (calculated as the average proportion of atretic cells in fish displaying atresia) of atresia were low in prespawning fish, but high from onset of spawning throughout the spawning period. Atretic oocytes categorized as in early alpha and in late alpha state occurred irrespective of maturity stage from late prespawning individuals up to late spawning fish, showing that oocytes may become atretic throughout the spawning period. Observed prevalence of atresia throughout the spawning period was almost 40% with an intensity of c. 20%. This indicates extensive down-regulation, i.e. considerably lower realized (number of eggs spawned) v. potential fecundity (number of developing oocytes), suggesting significant variability in reproductive potential. The extent of fecundity regulation in relation to fish condition (Fulton's condition factor) is discussed, suggesting an association between levels of atresia and fish condition.

This study investigated the biochemical composition of ovarian fluid and its effect on the fertilization capacity of turbot Scophthalmus maximus during the spawning season. The fertilization rate and pH of ovarian fluid varied throughout the spawning season, with the highest values recorded at the mid-season. Positive correlations were found between the fertilization rate and the ovarian fluid pH. The composition of major inorganic ions (Na(+) , K(+) , Ca(2+) and Cl(-) ) showed no significant changes during the spawning season. Alkaline phosphatase (AKP) activity was significantly higher during mid-season than other seasons. The lowest levels of protein, acid phosphatase (ACP) and aspartate aminotransferase (AAT) were in the ovarian fluid released at the mid-season. Moreover, significant relationships were observed between the fertilization rate and the levels of protein, ACP, AKP and AAT. These observations suggest that the biochemical profile of ovarian fluid affects the insemination microenvironment as well as the fertilization capacity of S. maximus eggs. Determination of such profiles may prove to be a useful strategy to improve S. maximus breeding techniques.

The translationally controlled tumour protein (TCTP) of turbot Scophthalmus maximus (SmTCTP) contains only one cysteine (Cys¹⁷⁰) at the C-terminal end. The biological role of this C-terminal Cys¹⁷⁰ in the antioxidation and growth-promoting functions of SmTCTP was examined by site-directed mutation of C170A (Cys¹⁷⁰ →Ala¹⁷⁰). It was found that C170A mutation not only obviously decreased the antioxidation capacity of the mutant-smtctp-transformed bacteria exposed to 0·22 mM hydrogen peroxide, but also significantly interrupted the normal growth and survival of the mutant-smtctp-transformed bacteria and flounder Paralichthys olivaceus gill (FG) cells, indicating a key role played by Cys¹⁷⁰ in the antioxidation and growth-promoting functions of SmTCTP. This study also suggested that the self-dimerization or dimerization with other interacting proteins is critical to the growth-promoting function of SmTCTP.

In March 2006, a 360 km2 no-take zone (NTZ) was established north of Gotland in the central Baltic Sea, with the purpose to scientifically evaluate the effects of a fishing ban on flatfish populations. A monitoring programme was set up to study the populations in the NTZ and in a reference area east of Gotland where the fishing pressure was high. The programme included fishing with multimesh survey nets, modelling of potential larval export and estimation of fish consumption by large marine predators. Overall, the results showed a clear positive effect of the NTZ on turbot Scophthalmus maximus, with higher densities in the closed area compared with the fished area and also higher densities after closure compared with before. The NTZ also had older individuals and a more even sex ratio. This, in combination with a high potential for larval export from the NTZ to Gotland, shows that the marine reserve may be important for maintaining a viable S. maximus stock at Gotland. Also, for flounder Platichthys flesus, the densities were higher in the NTZ compared to the reference area and there was a net larval export to the fished area. For both species, density-dependent growth was evident, with a lower length at age in the closed area. Potential predation by grey seal Halichoerus grypus and great cormorant Phalacrocorax carbo sinesis on flatfishes, that could hamper the evaluation of the marine reserve, was also addressed. Taken together, the results show that there are clear benefits of the fishing ban for both flatfish species within the NTZ, while the net effects on fisheries are difficult to quantify.

An ontogenetic sequence of Atlantic halibut Hippoglossus hippoglossus larvae, reared in intensive culture conditions, was cleared and stained and histologically processed to determine normal cranial chondrification for specimens ranging from 0 to 41 days post-hatch (dph). Twenty-six cranial cartilaginous structures were described, at daily intervals post-hatch. The ontogenetic trajectory, composed of alternating steps and thresholds, was interpreted as saltatory. In comparison with other flatfishes, H. hippoglossus exhibits delayed onset of chondrification. From 9 dph onwards, the ontogenetic trajectory resembles more than that of the turbot Psetta maxima than that of the common sole Solea solea or the summer flounder Paralichthys dentatus and winter flounder Pseudopleuronectes americanus. Hippoglossus hippoglossus with the gaping-jaw malformation, common in intensively cultured individuals of this species, were examined histologically. The reason larvae cannot close their mouth, as their yolk-sac resorbs, seems to be related to the fusion of the interhyal to the hyosymplectic and ceratohyal with which it is normally articulated.

Despite the fact that the stomach of turbot Psetta maxima is a curved tube that forms a half circle, it was demonstrated that gastric evacuation in this predatory flatfish fed natural prey closely followed the surface-dependent cylinder model developed from studies on gadoids with a straight stomach. Evacuation experiments were performed on two size groups of P. maxima fed sandeel Ammodytes tobianus as well as on P. maxima fed brown shrimp Crangon crangon at three different temperatures. This enabled the provision of a gastric evacuation model for studies on P. maxima, which takes into account the effects of the explanatory variables predator size, temperature, prey energy density and resistance of prey to the digestive processes in the stomach. Basically, the cylinder model predicts that a square-root function accurately describes gastric evacuation, which is inconsistent with the conclusion of a previous study on P. maxima that evacuation of A. tobianus is essentially linear with time. Use of the cylinder model to the values of the explanatory variables presented in the latter study, however, gave accurate predictions of the actually acquired evacuation data, which points to the generic value of the model.

International audience; A method for oxygenating and mixing suspensions of turbot Psetta maxima red blood cells (RBC) was tested in (31)P nuclear magnetic resonance (NMR) spectroscopy. In normoxia, the levels of inorganic phosphate (Pi) and nucleoside triphosphates (NTP) were stable up to 140 min and intracellular pH (pHi) was maintained and decreased oxygen partial pressure (P(O ( 2) )) from 30 to 15 and 600 Pa produced a significant fall in the intensity of NTP resonance, balanced by an increase in the Pi signal. Treatment of RBC with 0. 5 M isoproterenol during hypoxia exposure did not affect the pattern of changes in NTP or pHi induced by hypoxia and the effect was manifest only on Pi levels.

Amoebic gill diseases (AGD) caused primarily by the amphizoic Neoparamoeba spp. have been identified as significant to fish health in intensive aquaculture. These diseases have consequently received significant attention with regard to disease pathophysiology. Neoparamoeba perurans has been putatively identified as the aetiological agent in salmonids, with other species such as turbot Psetta maxima and sea bass Dicentrarchus labrax also affected. Similarly, Neoparamoeba spp. have also been identified in co-infections with other gill diseases in salmonids. While infection of the gills results in an acute multifocal hyperplastic host response, reduced gill surface area and increased mucous cell densities, ion regulation and respiration in terms of blood gasses are only marginally affected. This may be partially attributed to reserve respiratory capacity and a reduction in mucous viscosity allowing for a greater flushing of the gill, so reducing the gill mucus boundary layer. Clinical and acute infections result in significant cardiovascular compromise with increases in aortic blood pressure, and systemic vascular resistance in Atlantic salmon, Salmo salar, which are not seen in rainbow Oncorhynchus mykiss and brown trout Salmo trutta. Increases in vascular resistance appear to be due to vascular constriction potentially reducing blood flow to the heart in compromised fishes, the overall effect being to lead to a compensatory tissue remodelling and change in cardiac shape in chronically infected fishes. The combined effect of reduced gill surface area and cardiovascular compromise leads to a significant reduction in swimming performance and increases in the routine metabolic rate that lead to an increase in the overall metabolic cost of disease.

Freezing of adult flounder Platichthys flesus and turbot Psetta maxima for 24 h resulted in mean decrease of total length (LT) of 2·1 and 1·3% and a reduced body mass (M) of 3·1 and 1·3%, respectively. Increasing the freezing time to 25 days increased the shrinkage in turbot but not in flounder. The LT and MT conversion equations for the different species and time spent frozen are given. The differences between species could be explained by the higher volume to surface area relationship in turbot compared with flounder.

Food and feeding of juvenile turbot Scophthalmus maximus and flounder Pleuronectes flesus were studied in five nursery areas at Gotland, Central Baltic Sea, ICES SD 27 and SD 28. Ontogeny involved partitioning of available food resources. The food choice of turbot

The survival, growth, gonadal development and sex ratios of gynogenetic diploid turbot Scophthalmus maximus between 9 and 36 months of age were investigated. Gynogenesis was induced by activation of freshly collected eggs with diluted (1 : 10 with Ringer) and UVirradiated (30000 erg mm 2) sperm, followed by a cold shock at 1 to 0 C for 25 min, starting at 6 5 min after fertilization. The exclusive maternal inheritance of the resulting gynogenetic diploids was confirmed in all individuals used in the experiment by microsatellite markers. Mean total length and mass throughout the experiment was higher in the controls than in the gynogenetics. The gonadal development in gynogenetic males proceeded normally, and in gynogenetic females it was delayed during the first sexual maturation but was normal during the second one. The sex ratio was 1 male (M) : 1 female (F) in the controls, as expected, but 1 M : 3 F in the gynogenetics. In a second, independent experiment, carried out with fish originating from different broodstock, however, the sex ratio was 1 M : 1 F in the controls and 0 M : 1 F (i.e. 100% females) in the gynogenetics. Together, these results suggested that the main sex-determining genetic component in turbot fits well to female homogamety, in accordance with data from other flatfishes. These results indicate that it will be possible to obtain near or all-female turbot stocks through induced gynogenesis or by crosses involving hormonally sex-reversed fish., 2

Growth in volume of common dentex Dentex dentex and turbot Psetta maxima during larval development was studied by means of a quantitative histological method. A two-phase pattern of volume increase was recorded for both species, turbot volume being always higher than dentex volume. During the first phase, the increase was small but during the second phase volume rose sharply from 22 days post hatch (dph) and 17 dph onwards in dentex and turbot, respectively. In dentex, the specific growth rate (G) of the whole larva as well as that of all the structures studied (nervous tissue, trunk musculature, digestive tract, liver, pancreas, spleen and thymus) was always higher during the second phase, whereas in turbot, only total volume of the larva, trunk musculature and nervous tissue had a higher G during the same period. The pattern of allometric growth of digestive organs was similar for both species. These organs showed an initial positive allometric growth that later became near-isometric (digestive tract and liver) or negative (pancreas). In dentex, nervous tissue and trunk musculature showed near-isometry throughout the period studied. In turbot, nervous tissue exhibited negative allometry and trunk musculature changed from negative to positive allometry. In both species studied, the highest allometry coefficients were recorded for digestive organs before the larva switched to strict exotrophy. This would indicate the importance of the development of these organs for survival.

More than 800 000 reared juvenile turbot Psetta maxima were released in Danish waters during the period 1990 to 2002, as part of a stock enhancement programme. Performance of the released fish has been investigated using a combination of surveys and experimental studies. A total of 1278 wild fish and 439 alizarine marked released fish were caught in 16 surveys from 1993 to 1997. Recaptures were made over a period of c. 2 years after release. Results of growth analysis and measurements of nutritional status (condition factor, liver index, liver dry matter and otolith residual mass) showed only minor differences between wild and released fish. Growth rates of wild and of released fish were comparable to fish kept in the laboratory at the same temperature (specific length growth rate c. 0.5% day -1 at 75-185 mm total length and 12.5° C). The results indicated sufficient food in the release area, but significant differences in growth and in nutritional status, between years, were also observed. Nutritional status of both wild and released fish was comparable to fed fish in the experiments and significantly higher than in fish starved for > I week. The effects of the transport and release procedures used were examined by simulated releases in laboratory tanks. The fish were observed to be feeding and growing normally during the first week after simulated release, suggesting this species to be relatively robust and unaffected by the handling and release procedures.

The Mediterranean turbot Psetta maxima consists of two main genetically distinct lineages (western Mediterranean and ‘eastern secluded Mediterranean’ basins) as investigated by mitochondrial DNA analysis. Within the latter lineage, most haplotypes from the Sea of Azov were endemic and more than half of them derived from a single ancestral haplotype shared among all the eastern Mediterranean areas. There was no relation between morphotype variation in bony tubercles and mitochondrial genealogy.

Turbot juveniles were exposed to four ammonia concentrations (0·17 (L), 0·34 (M), 0·73 (MH) and 0·88 (H) mg l-1 NH3-N) for different exposure durations (28 days minimum to 84 days). Their physiological status and growth performances were compared to a control group (0·004 (C) mg l-1 NH3-N). No growth was observed in the H group, and by day 57, mass increase in the MH group was only 15% of that in group C. During the first month growth in the L group was similar to that in control group while it was lower (33%) in the M group; afterwards the L and M groups had a similar growth (half that of controls). Accumulation of total ammonia nitrogen (TA-N) in plasma was dependent on ambient ammonia concentrations. Plasma urea levels in ammonia-exposed fish were lower, similar or greater than in controls (depending on ammonia concentration or exposure duration). Osmolarity, Cl- and Na+ plasma concentrations were stable in the L and M groups. The increases in Na+, Cl-, K+ and total Ca concentrations observed by the end of the experiment in the H and MH groups suggest that fish failed to adapt. There was an initial rise in plasma cortisol in all ammonia-exposed groups followed by a return to basal level (1·7-4 ng ml-1) in the L and M groups. In group MH, plasma cortisol peaked at 42 ng ml-1 by day 14, and after a decline at c. 1 month (14 ng ml-1), it rose again.

A comparative study of blood oxygen binding and carrying capacities of turbot Scopthalmus, maximus and sea bass Dicentrarchus labrax, two fish species differing in their demand for oxygen, was carried out under three levels of chronic hypoxia (Po 2 =93. 65 and 40 mmHg) for 40 days. Blood O 2 affinity in normoxia was moderately high in both species (P 50 was c. 12-13 mmHg at pH 7.7). The Bohr factor was significantly lower in turbot (-0.52) than in sea bass (-0.85). In both species, blood O 2 affinity was not significantly affected by oxygen depletion whatever its level and duration. In turbot, however, P 50 appeared to slightly decrease at the two more severe levels of hypoxia. In both species, blood O 2 carrying capacity was not affected by hypoxia and remained twice as high in sea bass than in turbot.

When offered a size range of shrimp Crangon crangon, fewer naive reared turbot Scophthalmus maximus fed than did wild fish, and those that fed took smaller prey. Analysis of feeding behaviour indicated differences between wild and naive reared fish in the motivation to feed on novel prey and in prey recognition and capture efficiency. Feeding efficiency and motivation increased with experience and reared fish achieved the feeding rate of wild fish within the 9 days of the experiment. The ‘creep’ style of approach to shrimp was innate to naive reared turbot. A comparative feeding experiment confirmed that experienced reared fish ate more prey than naive reared fish. Naive reared turbot selected pellets and attacked stones preferentially to shrimp, in contrast to wild and experienced reared fish, which selected shrimp. Stones were attacked due to the memory of pellet-like visual characteristics and this behaviour persisted in some reared fish for at least 6 weeks, illustrating a cost of memory in which changing environmental conditions cause previously learnt information to become misleading. The experimental data also demonstrated that differences may arise in the prey and feeding behaviour of wild and naive reared fish, but that such differences diminish with experience.

Turbot Scophthalmus maximus maximum oxygen uptake following feeding and exhaustive exercise increased from 107 mg O2 kg−1 h−1 at 6° C to c. 218 mg O2 kg−1 h−1 at 18° C, then increased slightly from 18 to 22° C to 224 mg O2 kg−1 h−1. Standard oxygen uptake increased exponentially as a function of temperature from 11 mg O2 kg −1 h−1 at 6° C to 66 mg O2 kg−1 h−1 at 22° C. Gradual reduction in oxygen concentration to 87–90% air saturation at 6, 10. 18° C and 100% saturation) had little effect. Metabolic scope attained a maximum of 176 mg O2 kg−1 h−1 at 18° C. Interpolation of the results showed that this value changed little between 16 and 20° C. It is suggested that this temperature range is optimal for turbot of c. 500 g. A comparison with a previous study on feeding demand in intensive farming conditions showed a linear relationship between appetite and metabolic scope. It is concluded that the ability of a fish to supply energy (including the energy requirement of digestive metabolism) above a standard level is a limiting factor in the manifestation of its feeding demand.

Food conversion efficiency and growth of juvenile turbot Scophthalmus maximus, reared under temperatures of 14, 18 and 22° C, were found to be higher in one experiment (7-70 g fish) in a higher latitude (Norway) strain than lower latitude (France and Scotland) strains. In a second study (20-150 g fish), food intake, food conversion efficiency and growth were higher in higher latitude (Iceland and Norway) compared with a lower latitude (France) strain.

Turbot larvae were fed three different densities of rotifers (1000, 3000 and 7500 rotifers l -1 ) 1 ) with a low lipid level (

Pancreatic bile salt-dependent lipase (BSDL) was present with 0.5 μg BSDL larva -1 from hatching in turbot larvae. The enzyme content increased during the yolk sac phase to 1.1 μg BSDL larva -1 . This suggests that larval turbot are able to digest lipids from the start of exogenous feeding. The BSDL synthesis was stimulated first by food about 5 days after the onset of first feeding. The content per larva increased exponentially in fed larvae to 20 μg BSDL larva -1 on day 23 after hatching and decreased in starved larvae. In contrast, the specific content decreased during the first feeding phase, meaning that smaller larvae had a higher content of enzyme related to their biomass than did bigger larvae.

Growth performance of a high latitude (Norway) population of juvenile turbot Scophthalmus maximus was superior to that of two other lower latitude populations (Scotland, France) especially at 18° and 22° C. Overall these results lend some support to the hypothesis of countergradient variation in growth. The Norwegian population had the highest estimated temperature optimum for growth (T opt.G ± S.E.) (23.0 ± 0.9° C) and food conversion efficiency (T opt.Ec )(17.5 ± 0.3), followed by the French (T opt.G , 21.1 ± 1.0; T opt.Ec 16.7 ± 0.1) population, whereas the Scottish population had the lowest optimum (T opt.G , 19.6±0.6; T opt.Ec , 16.5 ± 0.1° C). These results have two major implications: firstly, for turbot culture, particularly in selection work focusing on growth performance; secondly, if countergradient variation in growth performance takes place within a species one cannot assume automatically that one set of physiological parameters, in this case growth-related parameters, is satisfactory to predict growth for a species throughout its range as different populations might show a difference in response towards different physiological parameters.

Exposure of adolescent turbot Scophthalmus maximus to low rations during vitellogenesis, covering the 4 months immediately prior to spawning, led to a drop of 70% in mean ovary weight, and was associated with poor growth of the vitellogenic oocytes or, in a third of cases, the absence of vitellogenic oocytes. Exposure to low rations during the recruitment of vitellogenic oocytes, 4 8 months prior to spawning, produced a more variable response. A few of these females produced well-developed vitellogenic oocytes, but in most, vitellogenic oocytes were absent or only poorly developed and in some cases there was a high incidence of atresia, Exposure to intermediate rations throughout oocyte recruitment and vitellogenesis also depressed oocyte development. In maturing females the number of non-atretic vitellogenic oocytes (relative potential fecundity) averaged 998 per g of somatic body weight of fish at the start of the spawning season in mid-June. Maturation of the males was not significantly influenced by the dietary regime. The small size of the testes and the low levels of milt production, even in fish fed high rations immediately prior to spawning, showed that reproductive investment in males was much lower than in females.

The development of a recombinant turbot somatotropin (rtuGH) in Escherichia coli and an homologous antibody in rabbit has produced a sensitive and specific RIA allowing the measurement of blood levels and pituitary contents of tuGH and their responses to environmental change. Turbot IGF-1 (insulin growth factors) and IGF type 1 receptor (R) cDNA have also been cloned and sequenced. The deduced IGF-1 R primary sequence contains all the topological features characteristics of mammalian IGF-1 R and the turbot R appears highly conserved, compared to its mammalian counterpart, particularly within the catalytic domain. IGF-1 R mRNA polyadenylation status varies during development with polyadenylation occuring in oocytes and early stage larvae. It disappears in later stages and in adult somatic tissues, indicating that IGF-1 R mRNA undergoes complex post-transcriptional regulation. Thyroid hormones (TH, both T 3 and T 4 ), are also involved in growth control, but are less affected by environmental changes than GH, except in very extreme conditions. The thyroid stimulating hormone (TSH) β-subunit cDNA fragment has also been cloned and sequenced. mRNA expression levels have been quantified under conditions where blood circulating TH were modified by dietary treatments or hormone supplementation. Specific growth rate in turbot is related to food intake and is doubled with a temperature increase from 8 to 20° C. Lipid storage by contrast was higher at low temperatures. Over a long time course, growth rate in turbot can be improved in brackish water (8-20 psu) but it cannot adapt to very low salinities. Photoperiod appears to have little influence on growth when feeding is unrestricted. Under intensive farming conditions, ammonia, the major nitrogen waste product in teleosts. may increase to levels that can reduce growth. Safe ambient levels for growth are

The drinking rate of turbot larvae increased from 14 to 120 nl larva-1 h-1 from day 2 to 11 after hatching, which gave a slightly increased specific drinking rate (calculated per biomass) from day 2 to 7 (0·8–1·9 nl μg carbon-1 h-1. The clearance rate of both algae and bacteria was 10–100 times higher than the drinking rate, which indicated that the larvae had an active uptake of both algae and bacteria. On day 2 and 4 after hatching highest clearance rate was observed for Tetraselmis sp. On day 6 about the same clearance rate was observed for bacteria, Isochrysis galbana and Tetraselmis sp. Until day 4 the turbot larvae had a higher ingestion rate of Tetraselmis sp. than of I. galbana, whereas on day 6 the rates were similar (28–41 ng carbon larvae-1 h-1). The assimilation efficiency was somewhat higher for I. galbana than for Tetraselmis sp., and on day 6 the assimilated algae constituted 1·5 and 0·9% of the larval biomass for I. galbana and Tetraselmis sp., respectively.

Causes of size variation in a population of juvenile turbot were studied using an individual based model (IBM). Each simulation started with 800 (divided into eight groups of 100 each) 120-day-old (posthatch) juveniles and was run for 140 days, and the data gained from model simulations compared directly with the result of a laboratory study with size-graded turbot. Stochastic growth with memory, which was included in the models as an individual genetical growth rate variation, is important in explaining size variation, and the combination between individual genetic growth rate and social interactions related to size-dependent hierarchies also contributes to size variation. The use of size-dependent growth rate alone fails to explain size variation, and is of little value in predicting size variation in turbot culture. Further, the results indicate formation of different types of size hierarchies for different sizes of juvenile turbot.

Turbot Scophthalmus maximus is the focus of a rapidly expanding aquaculture industry, while at the same time wild catches appear to be in decline. As a preliminary investigation into the effects of hatchery rearing, genetic variation at three polymorphic microsatellite loci was assessed in two wild populations and two farmed strains of turbot, from Ireland and Norway. Although a considerable loss of rare alleles was observed in the Irish farmed strain, no statistically significant reductions were found in mean heterozygosity or allelic diversity in farmed strains compared to wild populations. Significant genetic heterogeneity was found between wild and farmed samples from each country but not between the two wild populations. Genetic differentiation between the farmed strains was presumed to be caused by drift in the hatcheries. The utility of these particular microsatellite loci in comparing these samples and the importance of molecular genetic testing of farmed strains is stressed.

The deleterious effect of the ageing phenomenon of turbot spermatozoa was investigated in relation to the sampling date. Spermatozoa with a low or highly condensed chromatin and a middle piece containing numerous or a few vesicles were observed simultaneously 80 and 47 days before the beginning of the spawning period of the females. The middle piece of spermatozoa contained few vesicles, 39 days after the end of the reproductive period. At the same date, some spermatozoa appeared in which the plasma membrane was broken. Sperm motility, assessed just after collection in terms of arbitrary motility scores from 0 to 5, was significantly increased both at 10 and 60 s post-activation, for samples collected 18 days after, 25 days before and 9 days after the beginning of the spawning period of the females, respectively compared to samples collected 6 days before, 55 days after and 88 days after the end of this period. A lower short-term storage capacity was recorded at 10 and 60 s post-activation for sperm samples collected 6 days before and 88 days after the end of the reproductive period, respectively compared to 18 days and 9 days after the beginning of the spawning period. At 60 s post-activation, a higher motility of thawed spermatozoa was observed for samples collected 5 days before the beginning of the spawning period (motility recovery index: 86.4 ± 19.4%) compared to 71 days after the end of this period (55.0 ± 12.0%). The fertilizing capacity of sperm samples collected 61 days after the end of the spawning season (66.1 ± 14.6%) was significantly lower than that recorded for samples collected 34 days after the beginning of the spawning period (75.2±9.6%). On the contrary, there was no significant decrease in endogenous ATP content (31 days after the beginning of the spawning period, 14.53 ± 0.84; 48 days after the end of this period, 10.75 ± 5.26 nmol 10− 8 spermatozoa). Furthermore, sperm concentration significantly increased between the same dates (respectively 3.3 ± 0.8–9.4±4.8×109 spermatozoa ml−1).

Ichthyophonus sp. is reported for the first time in Mugil capito (thinlip grey mullet) and Li a saliens (leaping grey mullet). The fungus was also found in L. aurata (golden grey mullet), Dicentrarchus labrax (sea bass), Sparus aurata (gilthead sea bream) and Scophthalmus maximus (turbot), whereas Mugil cephalus (grey mullet) was not parasitized. In fish sampled periodically, the highest prevalences were observed in sea bass and the lowest in turbot. Among the fish sampled occasionally, the fungus was found associated to an epizootic in thinlip grey mullet. Ichthyophonus was never found in fish weighing

Reared 0-group turbot originating from Irish Sea stock were heavier at a given length than wild fish due to a greater body width and body depth. A very high incidence (96%) of abnormal lateral line canals was noted in the reared fish. It is suggested that these morphometric and morphological differences developed during the larval stage.

The appearance of large supranuclear vacuoles in the enterocytes of 1- and 4-days post-hatch larvae of turbot and herring, respectively, revealed by the pinocytotic uptake of a fluorescent marker (FITCdextran), indicates a potential for the absorption of dissolved nutrients by the endotrophic stages of marine fish larvae. Ingestion of algal cells by turbot larvae was observed soon after hatching, but low level pinocytotic absorption of algal material was first seen during the second day. More extensive lysis of algal cells and pinocytotic absorption occurred 24 h later. Although lysis of I. galbana was shown to occur at low external osmolarities, it is unlikely that sufficiently low osmolarities present in the hind gut of turbot larvae explained the observed rupture of algae. Other mechanisms for digestion are discussed. In newly hatched herring larvae, algal cells were unable to pass beyond the constriction in the mid-gut caused by the yolk sac. When algal cells were eventually seen to pass into the hind gut, there was no evidence of algal digestion or absorption throughout the remaining endotrophic and early exotrophic stages of herring larvae, although pinocytosis was observed to occur mid-way through the endotrophic stage.

Turbot larvae (24–590 degree C days; 2–32 days post-hatch) and juveniles (1345 degree C days; 98 days post-hatch), were exposed for 6 h to 25, 33 and 50% water-soluble fraction (WSF) of crude oil in either static or flow-through test systems. Larvae showed generalized primary endocrine responses, identified by elevated whole body cortisol content from as early as 2 days post-hatch. In older larvae and juveniles, the response was related to the WSF concentration. This dose-response relationship was not apparent in younger and yolk-sac larvae. Whole body thyroxine content of turbot larvae exposed to the WSF of crude oil was increased, but triiodothyronine content remained stable. Aromatic hydrocarbon concentrations [benzene, toluene, ethylbenzene and xylene (BTEX) and naphthalenes] remained constant during flow-through tests, but 65% of the initial level of BTEX hydrocarbons and 40% of the naphthalenes were lost during static exposures. Larval mortalities increased with exposure to an increasing concentration of crude oil WSF. Larval activity was significantly reduced even at the lowest WSF concentration.

Growth rate of individually tagged juvenile halibut was influenced significantly by the interaction of temperature and fish size. The results suggest an optimum temperature for growth of juvenile halibut in the size range 5–70 g between 12 and 15° C. Overall growth rate was highest at 13° C (1·62% day −1). At c. 5 g at the beginning of the experiment, fish at 16° C had the highest growth rate (3·2% day −1), but reduced this rate as they grew bigger. At 9 and 11°p C, growth rates were equal or only slightly lower during the later stages of the experiment, while the fish at 6° C showed significantly lower overall growth rate (0·87% day−1). Optimal temperature for growth decreased rapidly with increasing size, indicating an ontogenetic reduction in optimum temperature for growth. Moreover, a more flattened parabolic regression curve between growth and temperature as size increased indicated reduced temperature dependence with size. Although individual growth rates varied significantly at all times within the experimental temperatures, significant size rank correlations were maintained during the experiment. This indicated an early establishment of a stable size hierarchy within the fish groups. Haematocrit was highest at the highest temperature while Na+/K+-ATPase activity was inversely related to temperature. There was no difference in plasma Na+, Cl− and K+ concentrations among the temperature groups.

The effects of constant temperatures on growth, food efficiency, and physiological status were studied in four different batches of juvenile turbot. The growth responses were studied in three experiments lasting 70–85 days under 8–20° C thermal conditions. There was a positive correlation between growth and temperature from 8 to 17° C and a plateau was observed from 17 to 20° C. In fish fed to satiety, specific growth rate was positively correlated to the food intake, which was double at 20° C, compared with 8° C. Minor changes were observed in food efficiency. Body fat deposition decreased as temperature increased (25% lower at 20° C, compared with 8° C). Apparent food conversion, PER (protein efficiency ratio) and PUC (protein utilization coefficient) ranges were 0.8–0.9, 2.1–2.3 and 33–38% respectively. In 70–300 g fish, routine MO2 increased (2.5–6.5 μmol O2 h−1 g bw−1) with temperature up to 20° C, while larger turbot (500–600 g) appeared relatively thermo-independent, with a lower oxygen consumption (1.5 imol h−1 g−1). The average daily total ammonia nitrogen (TAN) and urea-N excretion per fish biomass was positively related to temperature. TAN was 30% lower at 8° C, compared with 20° C. Ingested nitrogen was mainly excreted under the final form of TAN, urea-N representing 26% of the total amount. A post-prandial peak in TAN and a delayed peak in urea-N nitrogen were observed. The hydromineral status [osmolarity, sodium, chloride and potassium blood plasma, gill (Na+-K+)-ATPase activity] of turbot was not affected by progressive changes in temperature during the acclimation period. Juvenile turbots show remarkable homeostatic capacities and so they have a relatively thermo-independent physiology within the range of temperature studied.

Turbot Scophthalmus maximus were cannulated via the afferent branchial artery and were either net-confined in sea water or in air for 9 min to monitor the stress response of a hatchery-reared marine flatfish. No mortality was observed. Aerial exposure appeared to mobilize plasma free fatty acids and stimulate the interrenal tissue but had no effect on circulating glucose or lactate levels. This pattern was qualitatively similar to that induced by net-confinement in sea water, although the magnitude and duration of the changes were more marked in the turbot handled in sea water. Aerial exposure had no effect on plasma osmolality, protein, or Cl− concentrations and only a minor effect on plasma Na+ concentrations, which was in sharp contrast to the ionoregulatory disturbance noted in turbot which were net-confined in sea water. However, plasma K+ and cortisol concentrations were similarly elevated by both handling procedures. The results from these experiments suggest that whilst similarities with the salmonid physiological stress response are apparent, the lack of a plasma glucose response may represent a fundamental difference in turbot stress physiology.

The effects of salinity changes (27, 19 and 10‰) on seawater-adapted juvenile turbot were studied on their plasma osmolarity and ion concentrations, on oxygen consumption, on gill Na+,K+-ATPase activity after 3 months and on growth parameters. All plasma concentrations (except chloride) were unchanged, suggesting that fish were well adapted to their environment. Oxygen consumption was significantly decreased in the 19 and 10‰ groups, where fish weighed significantly more 105 days after transfer than fish maintained in sea water. These results, and the fact that apparent food conversion rates were lower in a diluted environment, suggest that on a long term schedule growth conditions could be improved by adaptation to brackish waters (salinities between 10 and 19‰). The effects of transfer from sea water to 27, 19, 10 and 5‰ were also followed during the first 3 weeks. With salinity 10‰ a steady state was reached on day 21 with all plasma values within the same range. The significant differences observed in osmolarity, plasma ion concentrations and Na+,K+-ATPase activity 3 weeks after transfer of juveniles to 5‰ salinity, compared with transfers in higher salinities, suggest that there is a threshold of acclimation of turbot to a hypotonic environment.

In turbot larvae, Scophthalmus maximus, deprived of food for 24 h there was a significant increase in the specific surface of the epithelium and the corresponding microvillous border of the foregut accompanied by slight cellular degeneration. Following 48 h starvation Sarvae showed severe tissue degeneration in the foregut mucosa, progressing to extensive mucosal desquamation and cellular sloughing. Intracellular vacuolation of the epithelium and loss of microvilli was also extensive and the ability of the gut to absorb food must be severely impaired, with further starvation probably resulting in larval death. There was no evidence of parasitic infection in any of the larvae sampled and all observed alterations are attributable to food deprivation.

The efficiencies of water absorption from the guts of the larvae of herring (Clupea harengus L.) and turbot (Scophthalmus maximus L.) were estimated by two methods, The first method was based on the differences in the rates of accumulation, by drinking, and clearance from the gut of radiolabelled inert markers. The second method used the equilibrium level of radioactivity in the larvae to measure the concentration of the markers in the gut above background as water is absorbed from the gut. Water absorption efficiencies for herring larvae drinking sea water were estimated to be 77% using both methods. When external salinity was reduced to 50% sea water, drinking rates and water absorption efficiency in herring larvae fell substantially. Estimates of water absorption efficiency of seawater-adapted turbot larvae were similar (71–84%) to that of herring using both methods. Although temperature had a marked effect on both the rate of drinking and water absorption, there was no significant thermal effect on the efficiency of absorption from the guts of turbot larvae. The limitations of the techniques and the implications of the estimates in terms of water balance in fish larvae are discussed.

On sandy beach nursery grounds along the west coast of Ireland, 0 year-group turbot Scophthalmus maximus were found to consume six types of crustaceans, in addition to polychaetes. The 0 year-group brill Scophthalmus rhombus fed almost exclusively on mysids, even though nine taxonomic prey groups were identified in the sediment across the investigated beaches. Both species avoided non-motile organisms such as gastropods and bivalves, which were present in high abundances in the sediment and their growth and condition was not significantly related to the quantity or type of prey consumed, temperature or salinity. A high incidence of feeding was detected for both species over the duration of the study, suggesting that food was not limiting on west of Ireland nursery grounds. Temporal partitioning of settlement was detected between S. maximus and S. rhombus, indicating that inter-specific competition for food does not occur between these two flatfish species on west of Ireland nursery grounds.